No data available that match "Blotting, Southern"



*  Construction of the MsParA (Ms6939) knockout strain of | Open-i
... smegmatis and Southern blot assays.(A) Schematic representation of the recombination strategy for the ... D) Southern blot assays. A 300 bp probe corresponding to the sequences of the MsParA upstream genomic fragment of M. smegmatis ... D) Southern blot assays. A 300 bp probe corresponding to the sequences of the MsParA upstream genomic fragment of M. smegmatis ... 1B). Deletion of MsParA in the mutant strain was further confirmed by a Southern blot assay as shown in Figure 1D. Signal bands ...
  https://openi.nlm.nih.gov/detailedresult.php?img=PMC3366916_pone.0038276.g001&req=4
*  Glossary:Southern Blot
An assay that detects specific DNA molecules using a DNA or RNA probe with sequence similarity. Samples are subjected to electrophoresis on a slab gel. A replica of the gel is then made on a membrane by capillary transfer following denaturation. Specific DNA sequences are then detected on the membrane with a radioactively- or chemically-labeled probe. See the Figure from Alberts, et al., Molecular Biology of the Cell ...
  http://www.informatics.jax.org/glossary/southern_blot
*  Southern blotting - Biology-Online
And a western blot is the same but with protein (and Antibodies instead of a DNA probe). As a side note, Southern is the name ... DNA collected from a cell thought to contain the gene, then underwent the Southern blotting procedure. One of the fragments of ... Southern blotting. Discussion of all aspects of cellular structure, physiology and communication. ... I have heared of southern blotting for school but whats northern. eg how is it done, what is required? ...
  https://www.biology-online.org/biology-forum/viewtopic.php?t=2048
*  Southern Blot problems - Molecular Biology
Southern Blot problems - Help needed to fix southern problem!! (Oct/05/2006 ). Hi,. I have completed many many southern blots ... I blot using skimmed milk and dextran sulphate with 2 x SSPE and 1% SDS. I tried removing the dextran, it slightly improved but ...
  http://www.protocol-online.org/biology-forums/posts/20849.html
*  DIG southern blot - Molecular Biology - BioForum
DIG southern blot - posted in Molecular Biology: Please can someone give me some suggestions as to why my DIG SB is not working ... DIG southern blot. Started by cbrossin, Jun 03 2011 08:29 AM ... Then do a dot blot of serial dilutions of your DNA (no gel, no ... Then do a dot blot of serial dilutions of your DNA (no gel, no blotting) and hybridize with your probe to establish that the ... blotting) and hybridize with your probe to establish that the probe and DNA hybridize and can be detected. This also ...
  http://www.protocol-online.org/forums/topic/21471-dig-southern-blot/
*  Southern blot - Wikipedia
Other blotting methods (i.e., western blot, northern blot, eastern blot, southwestern blot) that employ similar principles, but ... acids Restriction fragment Genetic fingerprint Northern blot Western blot Eastern blot Southwestern blot Northwestern blot ... A Southern blot is a method used in molecular biology for detection of a specific DNA sequence in DNA samples. Southern ... Southern blots performed with restriction enzyme-digested genomic DNA may be used to determine the number of sequences (e.g., ...
  https://en.wikipedia.org/wiki/Southern_blot
*  DNA blot (Southern) - OpenWetWare
The DNA blot or Southern blot is a molecular biology method to probe the genome for the location of a DNA sequence. Genomic DNA ... DNA blot (Southern). From OpenWetWare. Revision as of 06:12, 16 April 2009 by Jakob Suckale. (talk , contribs) (lab-specific ... After size separation DNA pieces are blotted onto a membrane and then hybridised with a labelled nucleic acid probe. The ... Retrieved from "https://openwetware.org/mediawiki/index.php?title=DNA_blot_(Southern)&oldid=302398" ...
  https://openwetware.org/wiki/?title=DNA_blot_
*  PPT - SOUTHERN BLOTTING PowerPoint presentation | free to download - id: ad542-Y2IwN
The Southern blot is used to detect the presence of a particular piece of DNA in ... 6. Block with excess DNA. 7. Wash off ... Southern Blot - Southern Blot By: Jacqueline Jai Southern Blot Southern Blot-a piece nitrocellulose paper containing spots of ... Southern Blotting DNA Fingerprinting - Southern Blotting DNA Fingerprinting Southern Blot A Southern Blot identifies specific ... Southern, Northern and Western blotting - Southern, Northern and Western blotting * SOUTHERN BLOTTING The Random Hexamer ...
  http://www.powershow.com/view/ad542-Y2IwN/SOUTHERN_BLOTTING_powerpoint_ppt_presentation
*  Nachweis von TEL-Genrekombinationen mittels Southern Blot bei Kindern mit akuter lymphoblastischer Leukämie
The detection was done due to a new developed non-radioactive Southern blotting with a Digoxigenin marked template. We could ... Das in der vorliegenden Arbeit vorgestellte Verfahren der nicht-radioaktiven Southern Blot Hybridisierung unter Verwendung ...
  https://edoc.hu-berlin.de/handle/18452/15543?C=N
*  Blotting - Western / N / S > Southern Blotting Laboratory Product Directory and Equipment Reviews on...
Southern Blotting products in the SelectScience products and suppliers directory ... Read reviews and compare manufacturers of Blotting - Western / N / S > ... These Capillary Blotting systems offer a contained unit for southern and northern blotting. These units incorporate optimized ... Northern & Southern Blots Hybridization Agilent Technologies. MiracleHyb High-Performance Hybridization Solution QuikHyb ...
  http://www.selectscience.net/blotting---western-+-n-+-s/product-directory/southern-blotting/?catID=1535&u=F33BA311-66C8-47E5-AB3A-DAFF13E5CEFB
*  Southern blot
... 1. Photograph gel.. 2. Treat gel with 2 volumes of 0.2 N HCl and shake 15 min to depurinate DNA. Pour off ... 8. Blot membrane dry and place in glass cylinder preheated to 65oC. Add FBI hybridization buffer (1.5X SSPE, 10% PEG, 7% SDS) ...
  http://labs.icahn.mssm.edu/sorianolab/protocols/southern-blot/
*  Norther Blotting. Southern Blotting. Western Blotting. | Learning Material | Noodle
Southern Blotting. Western Blotting.. See what people are saying about Norther Blotting. Southern Blotting. Western Blotting.. ... Get an in-depth review and ask questions about Norther Blotting. ... Southern Blotting. Western Blotting.. Want more info about ... Southern Blotting. Western Blotting.? Get free advice from education experts and Noodle community members. ... Norther Blotting. Southern Blotting. Western Blotting.. http://www.dnaPot.com Follow us on twitter: http://www.twitter.com/#!/ ...
  https://www.noodle.com/learn/details/17540/norther-blotting-southern-blotting-western-blotting
*  Mutation detection by Southern blotting. - MyScienceWork
Mutation detection by Southern blotting.: Following the discovery of the structure of DNA in 1953, it became clear that ... Nevertheless, more than 30 years after its invention, the Southern blot remains a cornerstone of molecular biology. ... In 1975, Edward Southern published details of a new method for detecting DNA fragments based upon their specific sequence [ ... western and northern blot). The simplicity and effectiveness of the technique led to its universal acceptance as a standard ...
  https://www.mysciencework.com/publication/show/mutation-detection-southern-blotting-c5ff931a
*  The Use of Nested PCR and Southern Blotting to Confirm the Presence of Actin in Plasmodium yoelli | Biochemical Society...
The Use of Nested PCR and Southern Blotting to Confirm the Presence of Actin in Plasmodium yoelli. Ian D. Goodyer, Paul Towner ... The Use of Nested PCR and Southern Blotting to Confirm the Presence of Actin in Plasmodium yoelli ... The Use of Nested PCR and Southern Blotting to Confirm the Presence of Actin in Plasmodium yoelli ... The Use of Nested PCR and Southern Blotting to Confirm the Presence of Actin in Plasmodium yoelli ...
  http://www.biochemsoctrans.org/content/19/4/419S
*  Automated design of genomic Southern blot probes - Probe Design 1 - Genes2Cognition Neuroscience Research Programme
124 automated Southern blot designs Probe Design. Mouse Chr. Passed. Genomic Design Window (bases). Length Best Probe (bases). ... Designing a 'good' Southern blot probe for a particular gene or locus involves finding a stretch of DNA sequence at that locus ... full path to the southern_blot_design directory H. Program configuration files. Each of the five programmes requires a windows- ... Tiling algorithm for Southern blot probe design Given user-supplied chromosomal coordinates, and a desirable size range for the ...
  http://www.genes2cognition.org/software/southern_blot/
*  Southern and Northern Blotting | China-Mainland | Sigma-Aldrich
Northern Blotting Protocol. The protocol for Northern blotting is similar to that of Southern blotting. However, RNA sequences ... Southern blotting was introduced by Edwin Southern in 1975 as a method to detect specific sequences of DNA in DNA samples. The ... Southern and Northern blotting protocols involve the following major steps:. *Purification of DNA/RNA: Extract and purify the ... Place the blot in a film cassette lined with new saran wrap and carefully wrap the blot ensuring no air bubbles are trapped ...
  https://www.sigmaaldrich.com/china-mainland/zh/technical-documents/articles/biology/southern-and-northern-blotting.html
*  Will Southern s Patent Blot Out Affymetrix? | GenomeWeb
Struggling array-makers eagerly await the October 31 jury trial for Oxford Gene Technology's patent infringement suit against Affymetrix. Oxford is seeking $300 million in damages, an injunction, and/or royalties. Regarding its Delaware court date, Affy will say only that the patent in question is just one of many covering arrays.
  https://www.genomeweb.com/archive/will-southern-s-patent-blot-out-affymetrix
*  Southern Blotting Instrument Market Professional Survey Report 2017
Inquire for Southern Blotting Instrument Market Competitive Analysis, Trends and Forecast till 2022, with free sample copy of ... Southern Blotting Instrument industrySouthern Blotting Instrument industry analysisSouthern Blotting Instrument marketSouthern ... Blotting Instrument market 2017Southern Blotting Instrument market forecastSouthern Blotting Instrument market growthSouthern ... High Use of Southern Blotting Instrument in Automotive Industry Driving the Market Growth of Southern Blotting Instrument. ...
  http://newzy.net/southern-blotting-instrument-market/
*  Southern blotting
The technique of Southern blotting was developed by Sir Edwin Southern in 1973, and uses gel electrophoresis for the detection ... Tag Archives: Southern blotting. Preserving Science in the Archives (Part 1) 15 July 2013. 20th century, 21st century, ... These images represent a small selection of lab techniques such as Western and Southern blotting, and gel electrophoresis. ... There are also a number of dot blots, which is a method of applying proteins directly onto a membrane. Unlike Western blotting ...
  http://blogs.bodleian.ox.ac.uk/archivesandmanuscripts/tag/southern-blotting/
*  Next Biotechnology News | eBio World: Southern blot protocol
4. Southern blot. Perform a downward blot as described in the Red Book (Current Protocols). It works very well (complete ... Southern Blot (6) Spectroscopy (7) Stem cell (11) Theory of Ultracentrifugation (1) Top Stories (8) Type of Cancer (40) videos ... place light piece of plastic/glass on top of Southern to prevent evaporation, wait 1 hr (or more), dismantle Southern (gel ... Northern Blots (1) pcr (10) Pharmaceutical (1) Pharmaceutical Companies (3) Plant Biotech (16) PowerPoint (1) Projects(Doc) (5) ...
  http://www.ebioworld.com/2011/07/southern-blot-protocol.html
*  What is Southern Blotting? How it is used to identify DNA Sequences?
Southern Blotting is an elegant technique that can be used to identify the DNA sequences on restriction fragments separated by ... What is Southern Blotting? How it is used to identify Specific DNA Sequences?. March 29, 2016. Vinod Thakur Leave a comment ... Southern Blotting is an elegant technique that can be used to identify the locations of genes and other DNA sequences on ... Applications of Southern Blotting:. *An invaluable method for gene analysis and confirming DNA cloning results. ...
  https://sciencesamhita.com/what-is-southern-blotting-its-procedure-applications/
*  Northern and Southern blot analysis. (A) Northern blot | Open-i
A) Northern blot analysis of poly(A)+RNA isolated from the ears of Lewis, Brown Norway, Hooded Lister, Dark Agouti, and S ... Figure 9: Northern and Southern blot analysis. (A) Northern blot analysis of poly(A)+RNA isolated from the ears of Lewis, Brown ... Figure 9: Northern and Southern blot analysis. (A) Northern blot analysis of poly(A)+RNA isolated from the ears of Lewis, Brown ... C) Genomic Southern blot analysis of DNA from Sprague Dawley rats. The DNA was cleaved with three different restriction enzymes ...
  https://openi.nlm.nih.gov/detailedresult.php?img=PMC2196094_JEM.lutzelschwab9&req=4
*  Gene replacement strategy and Southern blot analysis of | Open-i
Gene replacement strategy and Southern blot analysis of progeny from heterozygous crosses. (a) The wild-type P-selectin allele ... A probe flanking the 3′-end of the vector was used for Southern blot analysis of ES cell clones and tail biopsies of the ... A probe flanking the 3′-end of the vector was used for Southern blot analysis of ES cell clones and tail biopsies of the ... b) Representative Southern blot analysis of tail biopsies of progeny from heterozygous crosses. Genomic DNA was digested with ...
  https://openi.nlm.nih.gov/detailedresult.php?img=PMC2132959_JCB9805113.f1b&req=4
*  Southern blot analysis of Mtb9.9 genes. Genomic DNA fro | Open-i
Southern blot analysis of Mtb9.9 genes. Genomic DNA from various mycobacterial strains was digested with PstI, separated by ... Southern blot analysis and examination of the Mtb genome sequence revealed a family of highly related genes. A T cell line from ... Southern blot analysis and examination of the Mtb genome sequence revealed a family of highly related genes. A T cell line from ... Figure 5: Southern blot analysis of Mtb9.9 genes. Genomic DNA from various mycobacterial strains was digested with PstI, ...
  https://openi.nlm.nih.gov/detailedresult.php?img=PMC2195809_JEM991786.f5&req=4
*  southern and northern on the same blot
... hrichter at ulys.unil.ch hrichter at ulys.unil.ch Thu Oct 20 18:42:11 EST 1994 * ... Hi out there, Are there some methods that would allow to have southern and northern on the same blot? I would like to quantify ...
  http://www.bio.net/bionet/mm/methods/1994-October/019884.html

No data available that match "Blotting, Southern"



(1/10141) Isolation of human transcripts expressed in hamster cells from YACs by cDNA representational difference analysis.

Gene isolation methods used during positional cloning rely on physical contigs consisting of bacterial artificial chromosomes, P1, or cosmid clones. However, in most instances, the initial framework for physical mapping consists of contigs of yeast artificial chromosome (YACs), large vectors that are suboptimal substrates for gene isolation. Here we report a strategy to identify gene sequences contained within a YAC by using cDNA representational difference analysis (RDA) to directly isolate transcripts expressed from the YAC in mammalian cells. The RDA tester cDNAs were generated from a previously reported hamster cell line derived by stable transfer of a 590-kb YAC (911D5) that expressed NPC1, the human gene responsible for Niemann-Pick type C (NP-C). The driver cDNAs were generated from a control hamster cell line that did not contain the YAC that expressed NPC1. Among the gene fragments obtained by RDA, NPC1 was the most abundant product. In addition, two non-NPC1 fragments were isolated that were mapped to and expressed from 911D5. One of these RDA gene fragments (7-R) spans more than one exon and has 98% sequence identity with a human cDNA clone reported previously as an expressed sequence tag (EST), but not mapped to a chromosomal region. The other fragment (2-R) that had no significant sequence similarities with known mammalian genes or ESTs, was further localized to the region of overlap between YACs 911D5 and 844E3. The latter YAC is part of a contig across the NP-C candidate region, but does not contain NPC1. This two-part approach in which stable YAC transfer is followed by cDNA RDA should be a useful adjunct strategy to expedite the cloning of human genes when a YAC contig is available across a candidate interval.  (+info)

(2/10141) Human papillomavirus DNA in adenosquamous carcinoma of the lung.

AIM: To investigate the presence of human papillomavirus (HPV) DNA in adenosquamous carcinoma of the lung--which is relatively common in Okinawa but not in mainland Japan--and examine its histological features. METHODS: Of 207 cases where primary lung cancers were surgically removed between January 1995 and June 1997 in Okinawa, 23 were adenosquamous carcinoma. HPV was detected by non-isotopic in situ hybridisation (NISH) and polymerase chain reaction (PCR) amplification with primers specific for E6 and E7 regions of the HPV genome. PCR products were analysed by Southern blotting. Immunohistochemical determination of high molecular weight cytokeratin (HMC) and involucrin was also carried out. RESULTS: 18 cases were positive for HPV DNA by PCR and NISH. HPV types 6, 11, 16, and 18 were found. Seven cases were dual positive for different types of HPV. Using NISH, HPV was also found in the squamous cell components and in neighbouring enlarged adenocarcinoma cells. The HMC and involucrin were demonstrated immunohistochemically in the same areas. CONCLUSIONS: HPV DNA was found in a high proportion (78.3%) of adenosquamous carcinomas in Okinawa, a region where HPV has previously been shown to be prevalent in squamous cell carcinoma of the lung. The adenocarcinoma cells adjacent to the squamous cell carcinoma component were enlarged and positive for HPV, HMC, and involucrin. This is thought to indicate the transition from adenocarcinoma to squamous cell carcinoma.  (+info)

(3/10141) The role of alternative splicing of the adhesion molecule, CD44, in lymphoid malignancy.

AIM: To investigate the expression of CD44 isoforms containing variant exon 6 (v6) in a well characterised cohort of patients with non-Hodgkin's lymphoma (NHL) and chronic lymphocytic leukaemia (CLL), and to correlate this with phenotype and disease course. METHODS: Cryostat sections of OCT embedded diagnostic nodal material from NHL patients and cryopreserved mononuclear preparations from CLL patients were used as sources of RNA. After reverse transcription, PCR was carried out with amplimers positioned at either side of the variant exon insertion site to amplify all possible CD44 isoforms. Those isoforms containing v6 were identified after Southern blotting and hybridisation with a radiolabelled oligonucleotide. RESULTS: Of 32 NHL samples analysed, 16 did not express CD44 isoforms containing v6, six expressed an isoform containing exon v6 alone, and 10 expressed v6 long isoforms which contained exon v6 in addition to other variant exons. These data did not correlate with lymphoma classification, disease staging, or the presence or absence of extranodal disease. However, those patients expressing v6 long CD44 isoforms had a worse overall survival than those that did not. The plateau of the survival curves was 50% compared with 82%. No v6 long isoforms were detected in the 21 CLL samples investigated. CONCLUSIONS: The expression of v6 long CD44 isoforms is associated with aggressive disease in NHL, independent of grade, stage, or presence of extranodal disease.  (+info)

(4/10141) Structure of cag pathogenicity island in Japanese Helicobacter pylori isolates.

BACKGROUND: cag pathogenicity island (PAI) is reported to be a major virulence factor of Helicobacter pylori. AIM: To characterise cagA and the cag PAI in Japanese H pylori strains. METHODS: H pylori isolates from Japanese patients were evaluated for CagA by immunoblot, for cagA transcription by northern blot, and for cagA and 13 other cag PAI genes by Southern blot. cagA negative strains from Western countries were also studied. Induction of interleukin-8 secretion from gastric epithelial cells was also investigated. RESULTS: All Japanese strains retained cagA. Fifty nine of 63 (94%) strains had all the cag PAI genes. In the remaining four, cag PAI was partially deleted, lacking cagA transcripts and not producing CagA protein. Details of the PAI of these strains were checked; three lacked cagB to cagQ (cagI) and continuously cagS to cag13 (cagII), and the remaining one lacked cagB to cag8. Western cagA negative strains completely lacked cag PAI including cagA. Nucleotide sequence analysis in one strain in which the cag PAI was partially deleted showed that the partial deletion contained 25 kb of cag PAI and the cagA promoter. Interleukin-8 induction was lower with the cag PAI partial deletion strains than with the intact ones. All Japanese cag PAI deleted strains were derived from patients with non-ulcer dyspepsia, whereas 41 of 59 (70%) CagA-producing strains were from patients with peptic ulcers or gastric cancer (p<0.05). CONCLUSIONS: Most Japanese H pylori strains had the intact cag PAI. However, some lacked most of the cag PAI in spite of the presence of cagA. Thus the presence of the cagA gene is not an invariable marker of cag PAI related virulence in Japanese strains.  (+info)

(5/10141) Comparison of Bombyx mori and Helicoverpa armigera cytoplasmic actin genes provides clues to the evolution of actin genes in insects.

The cytoplasmic actin genes BmA3 and BmA4 of Bombyx mori were found clustered in a single genomic clone in the same orientation. As a similar clustering of the two cytoplasmic actin genes Ha3a and Ha3b also occurs in another lepidopteran, Helicoverpa armigera, we analyzed the sequence of the pair of genes from each species. Due to the high conservation of cytoplasmic actins, the coding sequence of the four genes was easily aligned, allowing the detection of similarities in noncoding exon and intron sequences as well as in flanking sequences. All four genes exhibited a conserved intron inserted in codon 117, an original position not encountered in other species. It can thus be postulated that all of these genes derived from a common ancestral gene carrying this intron after a single event of insertion. The comparison of the four genes revealed that the genes of B. mori and H. armigera are related in two different ways: the coding sequence and the intron that interrupts it are more similar between paralogous genes within each species than between orthologous genes of the two species. In contrast, the other (noncoding) regions exhibited the greatest similarity between a gene of one species and a gene of the other species, defining two pairs of orthologous genes, BmA3 and HaA3a on one hand and BmA4 and HaA3b on the other. However, in each species, the very high similarities of the coding sequence and of the single intron that interrupts it strongly suggest that gene conversion events have homogenized this part of the sequence. As the divergence of the B. mori genes was higher than that of the H. armigera genes, we postulated that the gene conversion occurred earlier in the B. mori lineage. This leads us to hypothesize that gene conversion could also be responsible for the original transfer of the common intron to the second gene copy before the divergence of the B. mori and H. armigera lineages.  (+info)

(6/10141) Molecular cloning and characterization of three cDNAs encoding putative mitogen-activated protein kinase kinases (MAPKKs) in Arabidopsis thaliana.

We isolated three Arabidopsis thaliana cDNA clones (ATMKK3, ATMKK4 and ATMKK5) encoding protein kinases with extensive homology to the mitogen-activated protein kinase kinases (MAPKKs) of various organisms in the catalytic domain. ATMKK3 shows high homology (85% identity) to NPK2, a tobacco MAPKK homologue. ATMKK4 and 5 are closely related to each other (84% identity). Phylogenetic analysis showed that the plant MAPKKs constitute at least three subgroups. The recombinant ATMKK3 and ATMKK4 were expressed as a fusion protein with glutathione S-transferase (GST) in Escherichia coli. Affinity purified GST-ATMKK3 and GST-ATMKK4 proteins contained phosphorylation activity, which shows that both the ATMKK3 and ATMKK4 genes encode functional protein kinases. Northern blot analysis revealed that the ATMKK3 gene expressed in all the organs. The levels of ATMKK4 and 5 mRNAs were relatively higher in steins and leaves than in flowers and roots. We determined the map positions of the ATMKK3, 4 and 5 genes on Arabidopsis chromosomes by RFLP mapping using P1 genomic clones.  (+info)

(7/10141) Cloning, expression, and enzymatic characterization of Pseudomonas aeruginosa topoisomerase IV.

The topoisomerase IV subunit A gene, parC homolog, has been cloned and sequenced from Pseudomonas aeruginosa PAO1, with cDNA encoding the N-terminal region of Escherichia coli parC used as a probe. The homolog and its upstream gene were presumed to be parC and parE through sequence homology with the parC and parE genes of other organisms. The deduced amino acid sequence of ParC and ParE showed 33 and 32% identity with that of the P. aeruginosa DNA gyrase subunits, GyrA and GyrB, respectively, and 69 and 75% identity with that of E. coli ParC and ParE, respectively. The putative ParC and ParE proteins were overexpressed and separately purified by use of a fusion system with a maltose-binding protein, and their enzymatic properties were examined. The reconstituted enzyme had ATP-dependent decatenation activity, which is the main catalytic activity of bacterial topoisomerase IV, and relaxing activities but had no supercoiling activity. So, the cloned genes were identified as P. aeruginosa topoisomerase IV genes. The inhibitory effects of quinolones on the activities of topoisomerase IV and DNA gyrase were compared. The 50% inhibitory concentrations of quinolones for the decatenation activity of topoisomerase IV were from five to eight times higher than those for the supercoiling activities of P. aeruginosa DNA gyrase. These results confirmed that topoisomerase IV is less sensitive to fluoroquinolones than is DNA gyrase and may be a secondary target of new quinolones in wild-type P. aeruginosa.  (+info)

(8/10141) Reduced pyrazinamidase activity and the natural resistance of Mycobacterium kansasii to the antituberculosis drug pyrazinamide.

Pyrazinamide (PZA), an analog of nicotinamide, is a prodrug that requires conversion to the bactericidal compound pyrazinoic acid (POA) by the bacterial pyrazinamidase (PZase) activity of nicotinamidase to show activity against Mycobacterium tuberculosis. Mutations leading to a loss of PZase activity cause PZA resistance in M. tuberculosis. M. kansasii is naturally resistant to PZA and has reduced PZase activity along with an apparently detectable nicotinamidase activity. The role of the reduction in PZase activity in the natural PZA resistance of M. kansasii is unknown. The MICs of PZA and POA for M. kansasii were determined to be 500 and 125 micrograms/ml, respectively. Using [14C]PZA and [14C]nicotinamide, we found that M. kansasii had about 5-fold-less PZase activity and about 25-fold-less nicotinamidase activity than M. tuberculosis. The M. kansasii pncA gene was cloned on a 1.8-kb BamHI DNA fragment, using M. avium pncA probe. Sequence analysis showed that the M. kansasii pncA gene encoded a protein with homology to its counterparts from M. tuberculosis (69.9%), M. avium (65.6%), and Escherichia coli (28.5%). Transformation of naturally PZA-resistant M. bovis BCG with M. kansasii pncA conferred partial PZA susceptibility. Transformation of M. kansasii with M. avium pncA caused functional expression of PZase and high-level susceptibility to PZA, indicating that the natural PZA resistance in M. kansasii results from a reduced PZase activity. Like M. tuberculosis, M. kansasii accumulated POA in the cells at an acidic pH; however, due to its highly active POA efflux pump, the naturally PZA-resistant species M. smegmatis did not. These findings suggest the existence of a weak POA efflux mechanism in M. kansasii.  (+info)



  • gels
  • Make sure the gels boxes are very clean (any mineral oil will ruin the blot, and we have boxes set aside strictly for Southerns). (ebioworld.com)
  • probes
  • To address our own needs to regularly design Southern blot probes, automating this process, reducing the time needed to do this manually. (genes2cognition.org)
  • We calibrated the method using a set of 8 manually-designed mouse genomic probes (download here) that we have previously successfully employed for Southern blotting. (genes2cognition.org)
  • DNA microarrays operate by similar procedures to those used in the reverse northern blot, consisting of many DNA probes hybridized to a solid glass, plastic or silicon substrate which is probed with labeled RNA or cDNA. (wikipedia.org)
  • cDNA
  • Size markers are indicated at the left side of C. The blot was hybridized with the entire insert from the full-length cDNA clone for RMCP-8 and the blot was washed under medium stringency conditions (2 × SSC and 0.5% SDS at 65°C). (nih.gov)
  • sequence
  • If we're talking eukaryotic cells, then the answer might simply be that the RNA in the Northern blot was mRNA and had introns excised, therefore making it different in sequence than the DNA from whence it was transcribed. (biology-online.org)
  • samples
  • one should be a 0.6% gel for the Southern, which can use two combs for samples that are run halfway or a single comb for samples that can be run further (half way is fine to get a decent, publishable result on the Cambridge boxes). (ebioworld.com)
  • Bulked DNA samples can be analysed using Southern blotting. (wikipedia.org)
  • Methods
  • The names for other blotting methods may follow this convention, by analogy. (wikipedia.org)
  • These large regions were termed "ultra-long" telomeres in the literature when they were identified using southern blotting and "mega-telomeres" when identified by cytogenetic methods. (wikipedia.org)
  • similar
  • As a result, zoo blotting is used to detect similar or exact relationships between the DNA in question and other organisms. (wikipedia.org)
  • Definition
  • The key details related to Southern Blotting Instrument industry like the product definition, cost, variety of applications, demand and supply statistics are covered in this report. (newzy.net)
  • techniques
  • Although DNA Microarrays and newer next-generation techniques have generally supplanted reverse northern blotting, it is still utilized today and provides a relatively cheap and easy means of defining expression of large sets of genes. (wikipedia.org)
  • Type
  • This report divides the Southern Blotting Instrument market based on the key players, Type, Application and Regions are Mentioned Bellow. (newzy.net)
  • major
  • Competitive study of the major Southern Blotting Instrument players will help all the market players in analyzing the latest trends and business strategies. (newzy.net)
  • study
  • The study of emerging Southern Blotting Instrument market segments and the existing market segments will help the readers in planning the business strategies. (newzy.net)